Apparatus for vaccine development using ultrasound technology

ABSTRACT

Method and device for the creation of vaccines using ultrasonic waves, comprised of an ultrasound generator and a transducer to produce ultrasonic waves, is disclosed. The transducer has a specific ultrasound tip depending upon the type of delivery method utilized and depending on the shape of the vial containing the solution of the virus, bacterium, or other infectious agent. The apparatus delivers ultrasonic waves to solution either directly through the insertion of the ultrasound tip into the solution, through a coupling medium adjacent to the vial or near the vial, or through an air or gas medium. The ultrasound waves have the effect of destroying the viable virus, bacterium, or other infectious agent and of releasing the appropriate antigens, thus resulting in a vaccine for that virus, bacterium, or other infectious agent.

This application is a continuation of and claims benefit of U.S.application Ser. No. 11/393,180, filed Mar. 29, 2006, the teachings ofwhich are incorporated herein by reference.

BACKGROUND OF THE INVENTION

The present invention relates to the development of vaccines. Inparticular, the present invention relates to apparatus and methods fordeveloping vaccines using ultrasound technology.

Vaccine research and development has seen an increased level ofactivity, especially with the recent development of biodefenseinitiatives. The process of recombinant genetic engineering has provideda potential new approach to creating new and improved vaccines for thetreatment of disease. So far, this approach has met with limited successfor a variety of reasons, and thus many vaccines are still produced viatraditional methodologies.

Most classical vaccines are produced by one of two production methodsthat create either an inactivated (killed) or attenuated (live) vaccineproduct.

Inactivated vaccines (flu, cholera, hepatitis A) are produced by killingthe disease causing microorganism. A number of different methods ofinactivation can be used, including chemicals, irradiation, or heat.These vaccines are considered stable and relatively safe since theycannot revert to the virulent (disease-causing) form. The products oftendo not require refrigeration, a quality that makes them accessible anddesirable to domestic healthcare personnel as well as those indeveloping countries because they are practical for vaccinating peoplewho are in remote locations or involved in highly mobile activities(such as members of the armed force). However, most inactivated vaccinesproduce a relatively weak immune response and must be given more thanonce. A vaccine that requires multiple doses (boosters) may have alimited usefulness, especially in areas where people have limited accessto regular healthcare.

The second classical approach to the production of vaccines is anattenuated or live vaccine (measles, mumps, rubella). Thedisease-causing organism is grown under special laboratory conditionsthat cause it to loose its virulence or disease causing properties.Products prepared in this way require special handling and storage inorder to maintain their potency. These products produce both anti-bodymediated and cell-mediated immunity and generally they will only requireone booster dose.

While live vaccines do have some higher immune response advantages, thismethod of production has one large drawback. Because the organisms arestill living, it is their nature to change or mutate, causing theseproducts to have a remote possibility that the organisms may revert to avirulent form and potentially cause disease; thus, infection may occureither as a result of exposure while handling/processing the vaccine orafter administration of the vaccine. Therefore, these vaccines must becarefully tested and monitored. Patients who have compromised immunesystems are not usually administered live vaccines.

These two classical approaches to vaccine development and production notonly make up the majority of vaccines in use today, but these approachescontinue to be used in current vaccine development programs, includingthe development of vaccines for HIV/AIDS, newly identified variantstrains of Hepatitis, etc.

Alliger previously discussed using ultrasound technology to createvaccines in U.S. Pat. No. 5,582,829 (Alliger) and U.S. Pat. No.6,303,129 (Alliger). Alliger treats substantially viable cells, bacteriaor viruses (i.e. those that are intact and capable of functioning) withultrasound in order to make available antigens capable of inducing animmunogenic and/or therapeutic response. Specifically, the treatment ofcells and viruses with ultrasound is intended to deactivate thepotentially harmful cells and viruses and to also disperse the antigenspresent for use as a vaccine without further processing.

Alliger recommends that the procedure is conducted at room temperaturewhile maintaining the temperature of the sample containing the microbeagainst which a vaccine is developed between zero and 5 degrees Celsius.The minimization of heat is to prevent the denaturing of the antigens.Denaturing these antigens would limit their ability to produce aspecific immune response, thus diminishing the potential immunogeniceffect of the vaccine. The Alliger method is to deliver ultrasound at aspecific frequency, intensity, and duration in order to rupture anddestroy the viruses and bacteria within the sample through cavitation,to disperse the available antigens, and to do so without raising thetemperature of the viruses or bacteria to a level that would denaturethe antigens.

Alliger further states that the time must be sufficient to disrupt theviruses or cells so that no virulent cell structure remains to do this,Alliger states that one gram of cultured cells may generally requireabout 3 minutes of sonication.

As for sonicating the viruses and cells, Alliger delivered ultrasonicwaves to the microbe sample through a liquid medium at a frequency ofabout 20 kHz to about 40 kHz. He stated that above this frequency rangecavitation intensity is reduced considerably, even at high power inputs,so that cells or viruses may not be fully disintegrated. Alligerspecifically stated that the minimum intensity of the sonic waves shouldbe about 1 watt/sq. cm, and that the preferable intensity level at about20 kHz is 50 to 175 watts/sq. cm.

Alliger failed to mention the role of using different ultrasoundparameters and additional factors such as the volume of thesample/solution containing microorganisms and the geometrical shape ofthe ultrasound tip and vial/container to be used to achieve the mostefficient results in ultrasonic vaccine development. Because of theshortcomings of the classical approaches and Alliger's approach, thereis still a need for apparatus and methods that can produce inactivatedvaccines that can both produce a stronger immune response and that canproduce attenuated microorganisms for vaccine development incapable ofreverting back to a virulent strain.

SUMMARY OF THE INVENTION

The present invention is directed towards improvements of apparatusesand methods for the creation of vaccines using ultrasound wavespreviously researched and tested by the author of this patent in the1980's. Apparatus and methods in accordance with the present inventionmay meet the above-mentioned needs and also provide additionaladvantages and improvements that will be recognized by those skilled inthe art upon review of the present disclosure.

The present invention comprises an ultrasonic generator, an ultrasonictransducer, a sonication tip, and a vial or container of a solution thatcan be sonicated to create vaccines. The solution contained in the vialsis a mass of viruses, bacteria, or other infectious agents. The solutionis sonicated with ultrasound waves to destroy the viable infectiousvirus, bacteria, or infectious agent while also releasing theappropriate antigens, thus resulting in a vaccine for that virus,bacterium, or other infectious agent.

Ultrasound waves can be delivered to the solution either directlythrough the insertion of the ultrasound tip into the solution, through acoupling medium, or through an air/gas medium. The ultrasound tip thatis used can vary depending upon the type of delivery method chosen.There are three different types of recommended methods for sonicating asolution by the insertion of the ultrasound tip into the solutionitself. The first method uses a special shaped vial where the ultrasoundtip remains in the same position during the delivery of the ultrasoundenergy while the last two methods utilize movement of the ultrasound tipduring the sonication treatment.

There are also different types of recommended methods for sonicating thesolution through a coupling medium. There can be a medium placed betweenthe tip and the vial, there can be a liquid medium through which todeliver ultrasound waves, or the vial/container itself can be used as amedium if the tips is pressed up against the vial/container.

Based on the ultrasound intensity that is utilized, the sonication timeof the solution can vary. However, the intensity of the ultrasound wavescan be controlled through a variation in the ultrasound parameters suchas the frequency, the amplitude and the treatment time. The process mayrequire different intensity levels and ultrasound parameters based onthe specific type of virus, bacterium or other infectious agent used tocreate the vaccine and based on the volume of the solution containingmicrobes to be sonicated.

The invention is related to the apparatus and methods of deliveringultrasound energy to viruses, bacteria, or other infectious agents inorder to create a vaccine to treat the virus, bacterium, or infectiousagent.

One aspect of this invention may be to provide a method and device forthe creation of different vaccines.

Another aspect of the invention may be to provide a method and devicefor the creation of vaccines without the risk of toxicity that occurswith other chemical and temperature creation methods.

Another aspect of the invention may be to provide a method and devicefor the creation of high quality vaccines.

Another aspect of the invention may be to provide a method and devicefor the improvement of vaccine creation methods without usingtemperature or chemical influences.

Another aspect of the invention may be to provide a method and devicefor the creation of vaccines with a decreased production time.

Another aspect of the invention may be to provide a method and devicefor the continuous production of vaccines.

Another aspect of the invention may be to provide a method and devicefor the mass production of vaccines.

These and other aspects of the invention will become more apparent fromthe written descriptions and figures below.

BRIEF DESCRIPTION OF THE DRAWINGS

The present invention will be shown and described with reference to thedrawings of preferred embodiments and clearly understood in details.

FIG. 1 is a perspective view of an ultrasound vaccine development systemwhere the ultrasound tip is inserted into the solution.

FIG. 2 is a cross-sectional view of an ultrasound tip connected via acoupling medium to a vial.

FIG. 3 is a cross-sectional view of an ultrasound tip inserted into aliquid bath with a vial also inserted into the bath to deliverultrasound energy through the liquid to the vial.

FIG. 4 is a cross-sectional view of an ultrasound tip inserted into avial but located at a distance from the solution in the vial.

FIG. 5 are cross-sectional views of example ultrasound tips for use inthe ultrasound vaccine development system.

FIG. 6 are cross-sectional views of example different shaped vials foruse in the ultrasonic vaccine development system where the tip isinserted directly into the solution and sonicates the solution from aconstant position.

FIG. 7 is a cross-sectional view of recommended sonication methods touse with the ultrasound vaccine development system where the tip isinserted into the solution and moves during sonication.

FIG. 8 is a cross-sectional view of a production-line method to use withthe ultrasound vaccine development system.

FIG. 9 is a cross-sectional view of a carousel method to use with theultrasound vaccine development system.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is an apparatus and methods for the development ofvaccines using ultrasound technology. Preferred embodiments of thepresent invention in the context of an apparatus and methods areillustrated in the figures and described in detail below.

FIG. 1 illustrates the vaccine creation apparatus that has an ultrasonicgenerator 1, an ultrasonic transducer 2, a sonication tip 3, and a vial4 or other container in which a solution will be placed. The solution inthe vial or container is a mass of viruses, bacteria, or otherinfectious agents. The solution is sonicated with ultrasound waves todestroy the viable infectious virus, bacterium, or other infectiousagent while also releasing the appropriate antigens, thus resulting in avaccine against that virus, bacterium, or other infectious agent.Because the resulting vaccine is available for immediate use, theproduction time of vaccines developed through this method is lower thanthe production time of vaccines developed through classical methodsmentioned above. Ultrasound waves can be delivered to solution eitherdirectly through the insertion of the ultrasound tip into the solutionFIG. 1, through a coupling medium adjacent to the vial FIG. 2 or nearthe vial FIG. 3, or through the air or gas medium FIG. 4.

FIG. 5 shows examples of recommended ultrasound tips that can be useddepending on the type of delivery method. FIG. 5 a is a sphericalultrasound tip 13 and FIG. 5 b is a spherical ultrasound tip 14 thatcontains a central orifice 15. FIGS. 5 c/5 d/5 e/5 f show ultrasoundtips with a flat radiation surface. FIGS. 5 c and 5 e are ultrasoundtips 16 and 17 with a flat radiation surface, and FIGS. 5 d and 5 f areultrasound tips 17 and 20 with flat radiation surfaces and centralorifices 18 and 21. FIGS. 5 g and 5 h show ultrasound tips 22 and 23with a curved radiation surface. FIG. 5 e shows an ultrasound tip 23with a curved radiation surface and a central orifice 24. The centralorifices of the ultrasound tips shown in FIG. 5 can be used to deliversolution into a vial or container and/or can be used to providesonication during or after delivery of the solution.

FIG. 1 shows direct sonication where the ultrasound tip 3 is insertedinto the vial 4 and into the solution—the recommend tip 3 to use iseither a sphere FIGS. 5 a/5 b, a flat radiation surface FIGS. 5 c/5 d/5e/5 f, a rectangular prism (not shown), or another similar shape orcombination of shapes, with the sphere FIGS. 5 a/5 b as the preferredtip. The most preferred tip is the spherical tip 14 that contains acentral orifice 15; this is because the most preferred treatment methodinvolves the use of a spherical sonication tip where the solution isdelivered into the vial or container through the central orifice.

FIG. 2 shows delivery of ultrasound energy from an ultrasound tip 5through a coupling medium 6 such as liquid, gel, or the glass/plasticvial 7, where the tip 5 is pressed up against the vial 7 orcontainer—the recommended configuration of tip 5 is one that matches theshape of tip 5 to the geometric shape of the vial 7 or container. Forexample, if a spherical vial is to be sonicated, the recommend tip wouldbe a curved-shape tip (not shown) so that the tip would fit around theshape of the vial.

FIG. 3 shows delivery of ultrasound energy from an ultrasound tip 8through a liquid medium 9 where the ultrasound tip 8 is located at adistance from the vial 10—the recommended tip to use is a flat shapedtip FIG. 5 c/FIG. 5 d, with the preferred tip being a flat shaped tipwithout a central office as depicted in FIG. 5 c. For this method, theultrasound tip 8 is placed into the liquid medium 9 and deliversultrasound energy to the vial 10 through the liquid medium 9.

FIG. 4 shows delivery of ultrasound energy from an ultrasound tip 11 toa vial 12 through an air or gas medium—the recommended tip to use is aflat-shaped tip FIGS. 5 c/5 d/5 e/5 f, with the preferred tip eitherFIG. 5 c or 5 e. For this delivery method, the ultrasound tip 11 isinserted into the vial 12 but the tip 11 does not come into contact withthe solution in the vial 12.

FIG. 1 shows delivery of ultrasound energy where the ultrasound tip 3 isinserted into the vial 4 and into the solution—there are three differenttypes of recommended methods for this direct sonication. FIG. 6 showsthe first method that uses a special shaped vial where the ultrasoundtip remains in the same position during the delivery of the ultrasoundenergy, while FIG. 7 shows the last two methods that utilize movement ofthe ultrasound tip during the sonication treatment.

FIG. 6 shows the first method of direct sonication that uses both aspecial shaped vial 26, 28, or 30 and a corresponding ultrasound tip 25,27, or 29 that mirrors the shape of the vial 26, 28, or 30. There arethree different recommended shapes of vials 26, 28, or 30 to use with acorresponding ultrasound tip 25, 27, or 29: the three shapes are FIG. 6a a spherical vial 26, FIG. 6 b a rectangular vial 28, and FIG. 6 c acurved vial 30. With the spherical vial 26 shown in FIG. 6 a, aspherical shaped ultrasound tip 25 is inserted into the bottom of thevial 26. Because the ultrasound tip 25 mirrors the shape of the vial 26,there is an equidistant space between the ultrasound tip 25 and the vial26; this allows for the solution to be sonicated equally, thus resultingin an effective vaccine creation. This same concept of equal sonicationalso applies to the rectangular shaped vial 28 shown in FIG. 6 b. Arectangular-shaped ultrasound tip 27 that mirrors the shape of the vial28 is inserted into the solution therefore causing the solution to besonicated equally. Finally, the curved shaped ultrasound tip 29 shown inFIG. 6 c can be inserted into a curved shaped vial 30, thereforeallowing for equal sonication of the solution contained in the vial 30.The shapes of the vials 26, 28, or 30 contained in FIG. 6 are therecommend shapes, and the preferred shape is the spherical vial 26;other similar shapes or combinations of shapes of vials and ultrasoundtips can also be utilized.

FIG. 7 shows the second potential method of direct sonication where theultrasound tip 31 is inserted into the bottom of the vial 32 containingthe solution and then the tip rises in a continuous motion 33 as itdelivers ultrasonic energy. After the sonication begins, the ultrasoundtip 31 gradually rises to the top of the solution while deliveringultrasound energy. The ultrasound tip 31 stops its movement and stopsdelivering ultrasound energy after it reaches the top and the entiresolution has been sonicated. This movement during the delivery ofultrasound energy allows for equal sonication of the entire solution,which is effective because it ensures that the harmful cells and virusesare destroyed to prevent toxicity and that the antigens are released.This is more effective than inserting the tip to the bottom of a regularshaped vial and attempting to sonicate the entire solution from oneposition-delivering from one position results in varying sonicationbecause the distance of the solution to the ultrasound tip variesthroughout the vial.

FIG. 7 also shows the third potential method of direct sonication wherethe ultrasound tip 31 is inserted into the bottom of the vial 32containing solution and the tip 31 rises in a step-mode motion 34.Sonication occurs for a brief time and then stops. The ultrasound tip 31is moved slightly higher, and then sonication occurs again. Thisstep-delivery motion 34 is repeated until the tip 31 has moved to thetop and the entire solution has been sonicated. Similarly to thecontinuous movement delivery, this method allows for equal sonication ofthe entire solution. This distance between delivery steps in thisstep-mode delivery method can be of equal or varying distances.

FIG. 8 shows a cross-sectional view of a production-line sonicationmethod to use with the ultrasound vaccine development system. Vials 38move down the production line towards the ultrasound tip 35. Uponreaching the tip 35, the tip 35 moves down 37 into the vial 36 tosonicate the solution contained in the vial 36. After sonication theultrasound tip 35, moves back up 37 and waits until another vial 38moves to the ultrasound tip 37. This process is repeated to sonicatemultiple vials 38. There are multiple options in which the solution canbe inserted in the vials 38. Pre-filled vials 38 can be placed on theline, the ultrasound tip 37 can fill the vial 36 with the solutionthrough a central orifice (not shown) in the ultrasound tip 37, or therecan be a separate delivery mechanism/source or sources (not shown) thatcan fill the vials 38 as they move down the production line and towardsthe ultrasound tip 35. There are also multiple versions of the systemthat can be used—besides using different methods of filling the vialswith solution, one or more ultrasound tips can deliver ultrasonic energyto one or more vials at a time. Furthermore, different methods of directsonication where the ultrasound tip is inserted solution can also beused as described above.

FIG. 9 is a cross-sectional view of a carousel sonication method to usewith the ultrasound vaccine development system. Vials 42 are placed inthe carousel system and rotate around the carousel until they reach theultrasound tip 39. When the vial 40 reaches the ultrasound tip 39, thetip 39 moves down 41 into the vial 40 to sonicate the solution containedin the vial 40. After sonication, the ultrasound tip 39, moves back up41 and waits until another vial 42 moves to the ultrasound tip 39. Thisprocess is repeated to sonicate multiple vials 42. There are multipleoptions in which the solution can be inserted in the vials 38.Pre-filled vials 42 can be placed in the carousel, the ultrasound tip 39can fill the vial 40 with the solution through a central orifice (notshown) in the ultrasound tip 39, or there can be a separate deliverymechanism/source or sources (not shown) that can fill the vials 42 asthey rotate around the carousel and towards the ultrasound tip 39.Furthermore, different methods of direct sonication where the ultrasoundtip is inserted solution can also be used as described above. Theproduction line method and the carousel method are only recommendedsystems to sonicate vials of solution. Additional methods and systemscan be similarly effective.

Based on the ultrasound intensity that is utilized, the sonication timeof the solution can be from fractions of a second and above for bothpulse and continuous wave mode delivery. However, the intensity of theultrasound waves can be controlled through a variation in the ultrasoundparameters such as the frequency, the amplitude and the treatment time.The recommended frequency range for the ultrasound waves is 16 kHz to 20MHz, with the preferred frequency range of 30 kHz to 120 kHz, and themost preferred frequency value is 50 kHz. The amplitude of theultrasound waves can be 2 microns and above, with the recommendedamplitude to be in range of 3 microns to 250 microns, and the mostpreferred amplitude value is 80 microns. The recommended sonicationtreatment time is 5-10 seconds. The amount of solution in the vial is atleast 0.1 grams, and the preferred amount of solution is 5-10 grams.

The process may require different intensity levels and ultrasoundparameters based on the specific type of virus, bacterium or otherinfectious agent used to create the vaccine and based on the amount ofthe solution to be sonicated. For example, 5 ml of a solution can besonicated with an ultrasound frequency of 50 kHz, an amplitude of peakto peak of 50 microns, an intensity of about 1000 watts/cm², and thesonication time will take up to 10 seconds based on the type of virus,bacterium, etc solution. The longer the sonication time of the solution,the lower the level of intensity is required; the shorter the sonicationtime, the higher the level of intensity is required. The sonication ofthe solution can be conducted in different temperature environments, butthe preferred method is to use room temperature.

Although specific embodiments and methods of use have been illustratedand described herein it will be appreciated by those of ordinary skillin the art that any arrangement that is calculated to achieve the samepurpose may be substituted for the specific embodiments and methodsshown. It is to be understood that the above description is intended tobe illustrative and not restrictive. Combinations of the aboveembodiments and other embodiments as well as combinations of the abovemethods of use and other methods of use will be apparent to those havingskill in the art upon review of the present disclosure. The scope of thepresent invention should be determined with reference to the appendedclaims, along with the full scope of equivalents to which such claimsare entitled.

1. An device for creating vaccines comprising: an ultrasound generatordriving an ultrasound transducer; the ultrasound transducer having adistal end attached to an ultrasound tip; the ultrasound tip emittingultrasonic waves at a frequency and an amplitude to produce anultrasound dosage; a container for holding a solution with an infectiousagent; a central orifice within the ultrasound tip through whichsolution is added to the container; a means for controlling theultrasound dosage so that all portions of the solution achieveapproximately the same ultrasound dosage; the ultrasound dosagereleasing antigens from the infectious agent; and the ultrasound dosagedegrading the infectious agent viability.
 2. An device for creatingvaccines comprising: an ultrasound generator driving an ultrasoundtransducer; the ultrasound transducer having a distal end attached to anultrasound tip; the ultrasound tip emitting ultrasonic waves at afrequency and an amplitude to produce an ultrasound dosage; a containerfor holding a solution with an infectious agent; a central orificewithin the ultrasound tip through which solution is added to thecontainer; a means for controlling the ultrasound dosage so that allportions of the solution achieve approximately the same ultrasounddosage; the ultrasound dosage releasing antigens from the infectiousagent; and the ultrasound dosage inactivating the infectious agentviability.
 3. The device according to claim 1 having a carousel forholding a plurality of containers.
 4. The device according to claim 1having a substantially automated production process.
 5. The deviceaccording to claim 1 wherein the amplitude of the ultrasonic waves is atleast 2 microns.
 6. The device according to claim 1 wherein theamplitude of the ultrasonic waves is in the range of 3 to 250 microns.7. The device according to claim 1 wherein the amplitude of theultrasonic waves is approximately 80 microns.
 8. The device according toclaim 1 wherein the frequency of the ultrasonic waves is in the range of16 kHz to 20 MHz.
 9. The device according to claim 1 wherein thefrequency of the ultrasonic waves is in the range of 30 kHz to 120 kHz,10. The device according to claim 1 wherein the frequency of theultrasonic waves is in the range of approximately 50 kHz.
 11. The deviceaccording to claim 1 wherein the container may hold at least 0.1 gramsof solution.
 12. The device according to claim 1 wherein the containermay hold between 5 and 10 grams of solution.
 13. The device according toclaim 1 wherein the ultrasonic energy is delivered for a duration of atleast a fraction of a second.
 14. The device according to claim 1wherein the ultrasonic energy is delivered for a duration in the rangeof 5 to 10 seconds.
 15. The device according to claim 1 wherein at leasta portion of the ultrasound tip has a distal end shape selected from thegroup consisting of; a sphere, a rectangular prism, a flat surface, abulbous and a curved surface.
 16. The device according to claim 1wherein the shape of the ultrasound tip mirrors the shape of the vial inwhich it is inserted.
 17. The device according to claim 1 wherein thefrequency is selected from the group consisting of continuous, pulsedand modulated.
 18. The device according to claim 1 wherein a drivingwave form of the ultrasound transducer is selected from the groupconsisting of sinusoidal, rectangular, trapezoidal and triangular waveforms.